UNIVERSITI PUTRA MALAYSIA

PARVEEN DEVI A/P PATTIRAM

FSTM 2013 2

MICROENCAPSULATION AND FLAVOR STABILITY OF SPRAY-DRIED OLEORESIN FROM WATER DROPWORT (Oenanthe javanica) AND KACIP

FATIMAH (Labisia pumila)

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MICROENCAPSULATION AND FLAVOR STABILITY OF SPRAY-DRIED OLEORESIN FROM WATER DROPWORT (Oenanthe javanica) AND KACIP

FATIMAH (Labisia pumila)

By

PARVEEN DEVI A/P PATTIRAM

Thesis Submitted to the School of Graduate Studies, Universiti Putra Malaysia, in Fulfillment of the Requirement for the Degree of Master of Science

June 2013

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Abstract of thesis presented to the Senate of Universiti Putra Malaysia in fulfillment of the requirement for the degree of Master Science

MICROENCAPSULATION AND FLAVOR STABILITY OF SPRAY-DRIED OLEORESIN FROM WATER DROPWORT (Oenanthe javanica) AND KACIP

FATIMAH (Labisia pumila)

By

PARVEEN DEVI A/P PATTIRAM

June 2013

Chairman: Associate Professor Lasekan Olusegun Olaniyi, PhD

Faculty: Food Science and Technology

Water Dropwort (Oenanthe javanica) is a perennial herb with distinctive aroma and

taste. It is cultivated in marshy places of Asia and Australia. Besides that, “Kacip

Fatimah” (Labisia pumila) is a small herbaceous under shrub that roots and stems. There

were three objectives established in this study that are identification and sensory

characterization of compounds from water dropwort (Oenanthe javanica) and “Kacip

Fatimah” (Labisia pumila). Second objective is stability of aroma-active constituents in

oleoresin of Water dropwort (Oenanthe javanica) and “Kacip Fatimah” (Labisia pumila)

herbs during spray drying at 170±2oC and 180±2oC. Thirdly is to study the effect of

storage condition on the release flavors profiles of the encapsulated water dropwort

(Oenanthe javanica) and “Kacip Fatimah” (Labisia pumila) oleoresin. The first part in

this study was established by GC-MS and GC-O analysis to identify the chemical

constituents of the oleoresins from both water dropwort and “Kacip Fatimah” leaves. A

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total of 41 compounds were identified in Water dropwort. The major compound

obtained was richer in diterpenes such as incensole (9.04%). However, 33 compounds

were obtained from “Kacip Fatimah” and predominated with sesquiterpenes alcohol

such as T-cadinol (13.33%). Furthermore, stability of aroma-active constituents in

oleoresin of Water dropwort and “Kacip Fatimah” were successfully conducted by using

microencapsulation technique with a combination of gum Arabic (1.5%) and food grade

gelatin (2.5%) in the formulation of spray dried encapsulated leaves extracts whereby,

inlet temperatures of 170±2 oC and 180±2 oC with constant outlet temperature 84oC±4 oC

and the feed rate of 6 rpm were established for both leaves. At drying temperature 170±2

oC and 180±2 oC, a total of volatile compounds of water dropwort were reduced from 41

compounds to 9 and 6 compounds respectively. Incensole has the highest concentration

at these both drying temperatures (29.49% and 37.49%) respectively. Meanwhile, in

“Kacip Fatimah” also found reduction in number of volatile compounds from initially 33

compounds to 25 and 12 compounds respectively. T-cadinol has among the highest

concentration at these drying temperatures 170±2 oC (22.56%) and 180±2 oC (33.18%).

Finally, study the effect of storage condition on the release flavors profiles of the

encapsulated water dropwort (Oenanthe javanica) and “Kacip Fatimah” (Labisia

pumila) oleoresin was established. The microcapsules were stored under the same

temperature at 45±2 oC and 24 % RH for further studies. The microcapsules were

maintained for 30 days in a climate chamber. The numbers of volatile compounds in

microcapsules of water dropwort were reduced drastically after day 24. For instance, at

drying temperature 170±2 oC, encapsulated water dropwort has reduced to 2 compounds

as compared with day 6 (6 compounds). Encapsulated “Kacip Fatimah” also showed the

same reduction whereby, a total of 5 compounds were detected at 24 days of storage as

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compared to 25 compounds detected at 6 days storage. At 180±2 oC drying temperature,

encapsulated water dropwort and “Kacip Fatimah” showed the volatile compounds

reduced after 18 days of storage. A total of 4 and 3 compounds respectively were

detected as compare with 6 days of storage (5 and 2 compounds were identified)

respectively. Volatile compounds were no detected in both encapsulated water dropwort

and “Kacip Fatimah” at day 30 which is probably caused by the invisible cracks on the

surface of the microcapsules. Overall, particle size and emulsion size of microcapsules

also increased due to agglomeration occurs after storage. From this study, it can be

concluded that encapsulated water dropwort and “Kacip Fatimah”, incensole and T-

cadinol have the highest concentration during storage.

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Abstrak thesis yang dikemukakan kepada Senat Universiti Putra Malaysia sebagai memenuhi keperluan untuk ijazah Master Sains

PEMIKROKAPSULAN DAN KESTABILAN PERISA SEMBURAN-PENYALIRAN OLEORESIN WATER DROPWORT (Oenanthe javanica) DAN

KACIP FATIMAH (Labisia pumila)

Oleh

PARVEEN DEVI A/P PATTIRAM

Jun 2013

Pengerusi: Profesor Madya Lasekan Olusegun Olaniyi, PhD

Fakulti: Sains dan Teknologi Makanan

“Water dropwort” (Oenanthe javanica) ialah herba yang mempunyai aroma dan rasa

yang tersendiri. Kebanyakan herba ini terdapat di tempat yang paya di Negara Asia dan

Australia. Selain daripada itu, Kacip Fatimah (Labisia pumila) ialah herba yang kecil

yang tumbuh di bawah pokok renek dari akar dan batang. Terdapat tiga objektif

dijalankan dalam kajian ini iaitu Mengenalpasti dan pencirian deria sebatian daripada

“water dropwort” (Oenanthe javanica) and Kacip Fatimah (Labisia pumila). Objektif

kedua adalah kestabilan juzuk aroma-aktif dalam oleoresin daripada “water dropwort”

(Oenanthe javanica) and Kacip Fatimah (Labisia pumila) herba semasa pengeringan

semburan pada 170±2oC and 180±2oC. Ketiga adalah untuk mengkaji kesan keadaan

penyimpanan di atas rasa pembebasan profil terkandung “water dropwort” (Oenanthe

javanica) and Kacip Fatimah (Labisia pumila) oleoresin. Bahagian pertama dalam kajian

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ini telah menggunakan analisis GC-MS and GC-O untuk mengenalpasti juzuk kimia dan

aroma dari oleoresin dari kedua-dua daun tersebut. Jumlah sebanyak 41 sebatian meruap

telah dikenalpasti dalam “water dropwort”. Sebatian utama yang diperolehi adalah kaya

dengan diterpenes seperti incensole (0.95%). Walaubagaimanapun, 33 sebatian meruap

telah diperolehi dalam Kacip Fatimah dan dikuasai dengan alkohol sesquiterpenes

seperti T-cadinol (29.14%). Selanjutnya, kestabilan juzuk aroma-aktif dalam oleoresin

daripada “water dropwort” dan Kacip Fatimah telah berjaya dijalankan dengan

menggunakan teknik pemikrokapsulan dengan gabungan gam Arabik (1.5%) dan gelatin

gred makanan (2.5%) di dalam formulasi seburan kering terkandung daun dimana, suhu

dalaman 170±2 oC dan 180±2 oC dengan suhu luaran malar 84±4 oC dan kadar 6 rpm

telah ditubuhkan untuk kedua-dua daun. Pada suhu 170±2 oC dan 180±2 oC, jumlah

sebatian meruap dari “water dropwort” telah berkurang dari 41 kompaun ke 9 dan 6

kompaun masing-masing. Didapati incensole mempunyai kepekatan yang tertinggi

dalam kedua-dua suhu pengeringan (29.49% dan 37.49%) masing-masing. Sementara

itu, dalam Kacip Fatimah juga mendapati pengurangan dalam bilangan sebatian meruap

iaitu dari mulanya 33 kompaun ke 25 dan 12 kompaun masing-masing. T-cadinol

mempunyai antara kepekatan yang tertinggi di kedua-dua suhu pengeringan 170±2 oC

(22.56%) dan 180±2 oC (33.18%). Akhirnya, mengkaji kesan keadaan penyimpanan di

atas rasa pembebasan profil terkandung daripada “water dropwort” dan Kacip Fatimah.

Mikrokapsul telah disimpan di bawah suhu yang sama pada 45±2 oC dan 24% RH untuk

melanjutkan kajian. Mikrokapsul telah dikekalkan selama 30 hari di dalam ruang iklim.

Bilangan sebatian meruap dalam mikrokapsul “water dropwort” telah berkurang secara

drastik selepas hari ke-24. Sebagai contoh, pada suhu pengeringan 170±2 oC, seburan

kering terkandung “water dropwort” telah berkurang kepada 2 kompaun jika berbanding

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dengan hari ke-6 (6 kompaun). Seburan kering terkandung Kacip Fatimah” juga

menunjukkan pengurangan yang sama di mana sejumlah 5 kompaun telah dikesan pada

hari ke-24 jika dibandingkan dengan 25 kompaun dikesan pada penyimpanan 6 hari.

Pada suhu pengeringan 180±2 oC, seburan kering terkandung “water dropwort” dan

Kacip Fatimah telah menunjukkan sebatian meruap berkurang selepas 18 hari

pemyimpanan. Sebanyak 4 dan 3 kompoun masing-masing telah dikesan berbanding

dengan 6 hari penyimpanan (5 dan 2 kompoun telah dikenalpasti) masing-masing.

Sebatian meruap tidak dapat dikesan dalam kedua-dua “water dropwort” dan Kacip

Fatimah pada hari ke-30 yang mungkin disebabkan oleh keretakkan pada permukaan

mikrokapsul. Secara keseluruhannya, saiz zarah dan saiz emulsi mikrokapsul turut

meningkat akibat penumpuan berlaku selepas penyimpanan. Dalam kajian ini, dapat

disimpulkan bahawa “water dropwort” dan Kacip Fatimah, incensole dan T-cadinol

mempunyai kepekatan tertinggi semasa penyimpanan.

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ACKOWLEDGEMENTS

Praise to God for His help and guidance that finally I am able to complete my Master of

Science. First and foremost, I would like to extend my deepest gratitude to the entire

individual that has been involved in this research. First of all, a special thanks to my

supervisory committee Associate Professor Dr. Lasekan Olusegun Olaniyi, Professor Dr.

Tan Chin Ping and Professor Dr. Mohammad Zaidul Islam Sarker for their willingness

in overseeing the progress of my research work from its initial phases till the completion

of it. I do believe that all their advice and comments are for the benefit of producing the

best research work.

Secondly, to my mother, Deviki A/P Karam Singh and my brothers who have been a

constant source of support in emotional, moral and of course financial during my

postgraduate years, and this thesis would certainly not have existed without them. I also

dedicate this thesis to my beloved late father who made me interested in science

background and encouraged me to continue my studies.

I am also grateful to the staff of Faculty of Food Science and Technology, Universiti

Putra Malaysia for their kindness in handling their work. Last, but not the least to my lab

members, my close friends Shamala A/P Salvamani and Nurul Hanisah Binti Juhari and

to all my colleagues as well. Their view and tips are useful indeed.

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I certify that a Thesis Examination Committee has met on 13 June 2013 to conduct the final examination of Parveen Devi a/p Pattiram on her thesis entitled “Microencapsulation and flavor stability of spray-dried oleoresin from Water dropwort (Oenanthe javanica) and Kacip Fatimah (Labisia pumila)” in accordance with the Universities and University Colleges Act 1971 and the Constitution of the Universiti Putra Malaysia [P.U. (A) 106] 15 March 1998. The Committee recommends that the student be awarded the Master of Science.

Members of the Thesis Examination Committee were as follows:

Farinazleen Mohd Ghazali, PhD Title: Associate Professor Name of Faculty: Food Science and Technology Universiti Putra Malaysia (Chairman) Sharifah Kharidah Syed Muhamad, PhD Title: Associate Professor Name of Faculty: Food Science and Technology Universiti Putra Malaysia (Internal Examiner)

Abdul Karim Sabo Mohamed, PhD Title: Associate Professor Name of Faculty: Food Science and Technology Universiti Putra Malaysia (Internal Examiner) Zaibunnisa Abdul Haiyee, PhD Title: Senior Lecturer Name of Faculty: Fakulti Sains Gunaan Universiti Teknologi Mara Malaysia (External Examiner) _____________________ NORITAH OMAR, PhD Assoc. Professor and Deputy Dean School of Graduate Studies Universiti Putra Malaysia Date: 16 August 2013

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This thesis was submitted to the Senate of Universiti Putra Malaysia and has been accepted as fulfillment of the requirement for the degree of Master of Science. The members of the Supervisory Committee were as follows:

Lasekan Olusegun Olaniyi, PhD Associate Professor Faculty of Food Science and Technology Universiti Putra Malaysia (Chairman) Tan Chin Ping, PhD Professor Faculty of Food Science and Technology Universiti Putra Malaysia (Member) Mohammad Zaidul IslamSarker, PhD Professor Faculty of Pharmacy International Islamic University Malaysia (Member)

_____________________

BUJANG BIN KIM HUAT, PhD Professor and Dean School of Graduate Studies Universiti Putra Malaysia

Date:

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DECLARATION

I declare that the thesis is my original work expect for the quotation and citations which have been duly acknowledged. I also declare that it has not been previously, and is not concurrently, submitted for any other degree at Universiti Putra Malaysia or at any other institution.

___________________________ PARVEEN DEVI A/P PATTIRAM Date: 13 June 2013

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TABLE OF CONTENTS

Page ABSTRACT ii ABSTRAK v ACKNOWLEDGEMENT viii APPROVAL ix DECLARATION xi LIST OF TABLES xv LIST OF FIGURES xviii LIST OF APPENDICES xxi LIST OF ABBREVATIONS xxiv CHAPTER

1 INTRODUCTION 1

2 LITERATURE REVIEW 5 2.1 Water Dropwort (Oenanthe javanica) 5 2.2 Kacip Fatimah (Labisia pumila) 8 2.3 Oleoresin 11 2.4 Flavor 11 2.5 Microencapsulation 12 2.5.1 Classifications of microencapsulation 14 2.5.2 The uses of microencapsulation 16 2.5.3 Type of microencapsulation materials 16 2.6 Spray drying 17 2.7 Particle size 18 2.8 Scanning Electron Microscope (SEM) 19 2.9 Solvent extraction 20 2.10 Gas Chromatography (GC) 22 2.10.1 Gas Chromatography- Flame Ionization Detector (GC- FID)

22

2.10.2 Gas Chromatography-Mass Spectrometry (GC-MS) 25 2.10.3 Gas Chromatography-Olfactrometry (GC-O) 26 2.10.3.1 GC-O methodologies 27 2.10.3.2 Detection frequency 27 2.10.3.3 Direct intensity 28 2.10.3.4 Dilution to threshold 28 2.10.3.5 Human detector performance 29

3 IDENTIFICATION AND SENSORY CHARACTERIZATION OF COMPOUNDS FROM WATER DROPWORT (Oenanthe javanica) AND KACIP FATIMAH (Labisia pumila)

30 3.1 Introduction 31

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3.2 Materials and methods 31 3.2.1 Materials 31

3.2.2 Preparation of fresh herbs 31 3.2.3 Extraction of dried herbs 31 3.2.4 Gas Chromatography-Mass Spectrometry (GC-MS) analysis

32

3.2.5 Gas Chromatography-Olfactrometry (GC-O) analysis 33 3.2.6 Identification of compounds 33 3.3 Results and discussion 34 3.3.1 Quantification of the aroma-active constituents from Water dropwort (Oenanthe javanica) and Kacip Fatimah (Labisia pumila)

34

3.4 Conclusion 42

4 STABILITY OF AROMA-ACTIVE CONSTITUENTS IN OLEORESIN OF WATER DROPWORT (Oenanthe javanica) AND KACIP FATIMAH (Labisia pumila) HERBS DURING SPRAY DRIED AT 170oC AND 180oC

43 4.1 Introduction 43 4.2 Materials and methods 44 4.2.1 Encapsulation process 44 4.2.2 Moisture content determination 45 4.2.3 Quantification of encapsulated aroma-active constituents of herbs in the powder

46

4.2.3.1 Gas Chromatography-Mass Spectrometry (GC-MS) analysis

46

4.2.3.2 Gas Chromatography-Olfactometry (GC-O) analysis 4.2.4 Morphological characteristics by scanning electron microscope (SEM)

47

47 4.3 Results and discussion 4.3.1 Encapsulation process

48 48

4.3.2 Moisture content determination 4.3.3 Quantification of encapsulated aroma-active constituents of herbs 4.3.4 Morphological characteristic by scanning electron microscopy (SEM) 4.3.5 Particle size analysis 4.4 Conclusion

49

51

61 64 65

5 STUDY THE EFFECT OF STORAGE CONDITION ON THE RELEASE FLAVORS PROFILE OF THE ENCAPSULATED WATER DROPWORT (Oenanthe javanica) AND KACIP FATIMAH (Labisia pumila) OLEORESIN

67

5.1 Introduction 68 5.2 Materials and methods 69

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5.2.1 Release of flavor from the spray-dried powder at a constant temperature and humidity

69

5.2.2 Morphological characteristic by scanning electron microscopy (SEM)

69

5.2.3 Powder particle size analysis 69 5.2.4 Emulsion particle size 70 5.2.5 Aroma quantification of encapsulated powdered herbs during storage

70

5.2.6 Herbs’ flavors retention in stored powders 70 5.2.7 Release time-course of the encapsulated flavors 71 5.2.8 Statistical analysis 71 5.3 Results and discussion 72 5.3.3 Morphological characteristic by scanning electron microscopy (SEM) of the encapsulated Kacip Fatimah during the storage condition

80

5.3.4 Powder particle size analysis of storage encapsulated Water dropwort and Kacip Fatimah

87

5.3.5 Emulsion particle size of stored encapsulated Water dropwort and Kacip Fatimah

100

5.3.6 Moisture content analysis of storage encapsulated Water dropwort and Kacip Fatimah were collected after storage analysis

94

5.3.7 Retention of herbs’ flavors in the stored encapsulated powders Water dropwort and Kacip Fatimah

95

5.3.8 Quantification of encapsulated aroma-active constituents of Water dropwort (Oenanthe javanica) and Kacip Fatimah (Labisia pumila)that spray dried at 170±2oC and 180±2oC and stored for 30 days at constant temperature at 45±2oC and 25 % relative humidity

98

5.3.9 Release time course of the encapsulated Water dropwort (Oenanthe javanica) and Kacip Fatimah (Labisia pumila) during storage

119

5.4 Conclusion

121

6 SUMMARY, GENERAL CONCLUSION AND RECOMMENDATION FOR FUTURE RESEARCH

122

REFERENCES

125

APPENDICES 135 BIODATA OF STUDENT 149 LIST OF PUBLICATIONS 150